Assessment of Oxidative Stress Biomarkers in Rat Blood
Autor(a) principal: | |
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Data de Publicação: | 2023 |
Outros Autores: | , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.21769/BioProtoc.4626 http://hdl.handle.net/11449/248617 |
Resumo: | Redox status assessments are time-consuming, require a large volume of samples and great reagent amounts, and are not adequately described for methodological reproducibility. Here, the objective was to standardize redox balance determination, based on previously described spectrophotometric tests in pregnant rats, to improve precision, time dispensed, and the volume of samples and reagents, while maintaining accuracy and adequate cost benefits. This protocol summarizes oxidative stress markers, which focus on spectrophotometric tests for the assessment of thiobarbituric acid–reactive substances, reduced thiol groups, and hydrogen peroxide, as well as the antioxidant activity of superoxide dismutase, glutathione peroxidase, and catalase in washed erythrocyte and serum samples from full-term pregnant rats. For non-pregnant rats and other species, it is necessary to standardize these determinations, especially the sample volume. All measurements were normalized by the estimated protein concentrations in each sample. To establish optimum conditions for the reproducibility of the proposed methods, we describe all changes made in each assay’s steps based on the reference method reassessed for the new standardizations. Furthermore, the calculations of the concentrations or activities of each marker are presented. Thus, we demonstrate that the analysis of serum samples is easier and faster, but it is impossible to detect catalase activity. Furthermore, the proposed methods can be applied for redox balance determination, especially using smaller reagent amounts and lower sample volumes in lesser time without losing accuracy, as is required in obtaining samples during rat pregnancy. |
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Assessment of Oxidative Stress Biomarkers in Rat BloodBiomarkerLipid peroxidationOxidative stressPregnancyRedox statusRedox status assessments are time-consuming, require a large volume of samples and great reagent amounts, and are not adequately described for methodological reproducibility. Here, the objective was to standardize redox balance determination, based on previously described spectrophotometric tests in pregnant rats, to improve precision, time dispensed, and the volume of samples and reagents, while maintaining accuracy and adequate cost benefits. This protocol summarizes oxidative stress markers, which focus on spectrophotometric tests for the assessment of thiobarbituric acid–reactive substances, reduced thiol groups, and hydrogen peroxide, as well as the antioxidant activity of superoxide dismutase, glutathione peroxidase, and catalase in washed erythrocyte and serum samples from full-term pregnant rats. For non-pregnant rats and other species, it is necessary to standardize these determinations, especially the sample volume. All measurements were normalized by the estimated protein concentrations in each sample. To establish optimum conditions for the reproducibility of the proposed methods, we describe all changes made in each assay’s steps based on the reference method reassessed for the new standardizations. Furthermore, the calculations of the concentrations or activities of each marker are presented. Thus, we demonstrate that the analysis of serum samples is easier and faster, but it is impossible to detect catalase activity. Furthermore, the proposed methods can be applied for redox balance determination, especially using smaller reagent amounts and lower sample volumes in lesser time without losing accuracy, as is required in obtaining samples during rat pregnancy.Universidade Estadual PaulistaFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Laboratory of Experimental Research on Gynecology and Obstetrics Botucatu Medical School São Paulo State University (Unesp), São Paulo StateCenter of Natural and Human Sciences (CCNH) Federal University of ABC (UFABC), São Paulo StateLaboratory of System Physiology and Reproductive Toxicology Institute of Biological and Health Sciences Federal University of Mato Grosso (UFMT), Mato Grosso StateLaboratory of Experimental Research on Gynecology and Obstetrics Botucatu Medical School São Paulo State University (Unesp), São Paulo StateFAPESP: 2016/25207-5Universidade Estadual Paulista (UNESP)Universidade Federal do ABC (UFABC)Federal University of Mato Grosso (UFMT)Sinzato, Yuri K. [UNESP]Rodrigues, ThiagoCruz, Larissa L. [UNESP]Barco, Vinícius S. [UNESP]Souza, Maysa R. [UNESP]Volpato, Gustavo T.Damasceno, Débora C. [UNESP]2023-07-29T13:48:56Z2023-07-29T13:48:56Z2023-03-05info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlehttp://dx.doi.org/10.21769/BioProtoc.4626Bio-protocol, v. 13, n. 5, 2023.2331-8325http://hdl.handle.net/11449/24861710.21769/BioProtoc.46262-s2.0-85151483851Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengBio-protocolinfo:eu-repo/semantics/openAccess2024-08-16T14:12:50Zoai:repositorio.unesp.br:11449/248617Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-16T14:12:50Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Assessment of Oxidative Stress Biomarkers in Rat Blood |
title |
Assessment of Oxidative Stress Biomarkers in Rat Blood |
spellingShingle |
Assessment of Oxidative Stress Biomarkers in Rat Blood Sinzato, Yuri K. [UNESP] Biomarker Lipid peroxidation Oxidative stress Pregnancy Redox status |
title_short |
Assessment of Oxidative Stress Biomarkers in Rat Blood |
title_full |
Assessment of Oxidative Stress Biomarkers in Rat Blood |
title_fullStr |
Assessment of Oxidative Stress Biomarkers in Rat Blood |
title_full_unstemmed |
Assessment of Oxidative Stress Biomarkers in Rat Blood |
title_sort |
Assessment of Oxidative Stress Biomarkers in Rat Blood |
author |
Sinzato, Yuri K. [UNESP] |
author_facet |
Sinzato, Yuri K. [UNESP] Rodrigues, Thiago Cruz, Larissa L. [UNESP] Barco, Vinícius S. [UNESP] Souza, Maysa R. [UNESP] Volpato, Gustavo T. Damasceno, Débora C. [UNESP] |
author_role |
author |
author2 |
Rodrigues, Thiago Cruz, Larissa L. [UNESP] Barco, Vinícius S. [UNESP] Souza, Maysa R. [UNESP] Volpato, Gustavo T. Damasceno, Débora C. [UNESP] |
author2_role |
author author author author author author |
dc.contributor.none.fl_str_mv |
Universidade Estadual Paulista (UNESP) Universidade Federal do ABC (UFABC) Federal University of Mato Grosso (UFMT) |
dc.contributor.author.fl_str_mv |
Sinzato, Yuri K. [UNESP] Rodrigues, Thiago Cruz, Larissa L. [UNESP] Barco, Vinícius S. [UNESP] Souza, Maysa R. [UNESP] Volpato, Gustavo T. Damasceno, Débora C. [UNESP] |
dc.subject.por.fl_str_mv |
Biomarker Lipid peroxidation Oxidative stress Pregnancy Redox status |
topic |
Biomarker Lipid peroxidation Oxidative stress Pregnancy Redox status |
description |
Redox status assessments are time-consuming, require a large volume of samples and great reagent amounts, and are not adequately described for methodological reproducibility. Here, the objective was to standardize redox balance determination, based on previously described spectrophotometric tests in pregnant rats, to improve precision, time dispensed, and the volume of samples and reagents, while maintaining accuracy and adequate cost benefits. This protocol summarizes oxidative stress markers, which focus on spectrophotometric tests for the assessment of thiobarbituric acid–reactive substances, reduced thiol groups, and hydrogen peroxide, as well as the antioxidant activity of superoxide dismutase, glutathione peroxidase, and catalase in washed erythrocyte and serum samples from full-term pregnant rats. For non-pregnant rats and other species, it is necessary to standardize these determinations, especially the sample volume. All measurements were normalized by the estimated protein concentrations in each sample. To establish optimum conditions for the reproducibility of the proposed methods, we describe all changes made in each assay’s steps based on the reference method reassessed for the new standardizations. Furthermore, the calculations of the concentrations or activities of each marker are presented. Thus, we demonstrate that the analysis of serum samples is easier and faster, but it is impossible to detect catalase activity. Furthermore, the proposed methods can be applied for redox balance determination, especially using smaller reagent amounts and lower sample volumes in lesser time without losing accuracy, as is required in obtaining samples during rat pregnancy. |
publishDate |
2023 |
dc.date.none.fl_str_mv |
2023-07-29T13:48:56Z 2023-07-29T13:48:56Z 2023-03-05 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://dx.doi.org/10.21769/BioProtoc.4626 Bio-protocol, v. 13, n. 5, 2023. 2331-8325 http://hdl.handle.net/11449/248617 10.21769/BioProtoc.4626 2-s2.0-85151483851 |
url |
http://dx.doi.org/10.21769/BioProtoc.4626 http://hdl.handle.net/11449/248617 |
identifier_str_mv |
Bio-protocol, v. 13, n. 5, 2023. 2331-8325 10.21769/BioProtoc.4626 2-s2.0-85151483851 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Bio-protocol |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.source.none.fl_str_mv |
Scopus reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
|
_version_ |
1808128189560520704 |